Cefotaxime sodium

EUROPEAN PHARMACOPOEIA 8.0

Prepare each of 4 injection vials as shown in the table below :

Sample solution
(mL)
1.0

Solvent solution
(mL)
0

Water R
(mL)
4.0

1.0

1.0

3.0

1.0

2.0

2.0

1.0

3.0

1.0

Vial No.

C. 1-methyl-1H-tetrazole-5-thiol,

Static head-space conditions that may be used :

equilibration time : 15 min ;
D. (6R,7R)-7-amino-8-oxo-3-[(1H-1,2,3-triazol-4-ylsulfanyl)methyl]-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2 transfer-line temperature : 110 C.
carboxylic acid (7-TACA),
Temperature :
Column : 40 C for 10 min.
Heavy metals (2.4.8): maximum 5 ppm.
2.0 g complies with test C. Prepare the reference solution using
1 mL of lead standard solution (10 ppm Pb) R.
Water (2.5.12) : maximum 5.0 per cent, determined on 0.200 g. E. (6R,7R)-3-[(acetyloxy)methyl]-7-amino-8-oxo-5-thia-1azabicyclo[4.2.0]oct-2-ene-2-carboxylic acid (7-ACA),
Bacterial endotoxins (2.6.14) : less than 0.20 IU/mg, if
intended for use in the manufacture of parenteral preparations
without a further appropriate procedure for the removal of
bacterial endotoxins.
ASSAY
Liquid chromatography (2.2.29) as described in the test for
related substances with the following modications.
Injection : test solution (a) and reference solution (a).
System suitability : reference solution (a) :
repeatability : maximum relative standard deviation of
1.0 per cent after 6 injections.
Calculate the percentage content of cefoperazone sodium by
multiplying the percentage content of cefoperazone by 1.034.
STORAGE
In an airtight container, protected from light, at a temperature
of 2 C to 8 C. If the substance is sterile, store in a sterile,
airtight, tamper-proof container.

F. (6R,7S)-7-[[(2R)-2-[[(4-ethyl-2,3-dioxopiperazine-1-yl)carbonyl]amino]-2-(4-hydroxyphenyl)acetyl]amino]-3[[(1-methyl-1H-tetrazol-5-yl)sulfanyl]methyl]-8-oxo-5thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylic acid.
01/2008:0989

CEFOTAXIME SODIUM
Cefotaximum natricum

IMPURITIES

C16H16N5NaO7S2
[64485-93-4]

A. (5aR,6R)-6-[[(2R)-2-[[(4-ethyl-2,3-dioxopiperazin-1-yl)carbonyl]amino]-2-(4-hydroxyphenyl)acetyl]amino]5a,6-dihydro-3H,7H-azeto[2,1-b]furo[3,4-d][1,3]thiazine1,7(4H)-dione,

Mr 477.4

DEFINITION
Sodium (6R,7R)-3-[(acetyloxy)methyl]-7-[[(2Z)-2-(2aminothiazol-4-yl)-2-(methoxyimino)acetyl]amino]-8-oxo-5thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylate.
Semi-synthetic product derived from a fermentation product.
Content : 96.0 per cent to 102.0 per cent (anhydrous substance).
CHARACTERS
Appearance : white or slightly yellow powder, hygroscopic.
Solubility : freely soluble in water, sparingly soluble in
methanol.
IDENTIFICATION
A. Infrared absorption spectrophotometry (2.2.24).
Comparison : cefotaxime sodium CRS.
B. It gives reaction (a) of sodium (2.3.1).

B. (6R,7R)-7-[[(2R)-2-[[(4-ethyl-2,3-dioxopiperazin-1-yl)carbonyl]amino]-2-(4-hydroxyphenyl)acetyl]amino]-3-[(4- TESTS
methyl-5-thioxo-4,5-dihydro-1H-tetrazol-1-yl)methyl]-8Solution S. Dissolve 2.5 g in carbon dioxide-free water R and
oxo-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylic acid, dilute to 25.0 mL with the same solvent.
General Notices (1) apply to all monographs and other texts

1801

Cefotaxime sodium

EUROPEAN PHARMACOPOEIA 8.0

Appearance of solution. Solution S is clear (2.2.1). Add 1 mL

of glacial acetic acid R to 10 mL of solution S. The solution,
examined immediately, is clear.
pH (2.2.3): 4.5 to 6.5 for solution S.
Specic optical rotation (2.2.7) : + 58.0 to + 64.0 (anhydrous
substance).
Dissolve 0.100 g in water R and dilute to 10.0 mL with the
same solvent.
Absorbance (2.2.25) : maximum 0.40 at 430 nm for solution S.
Specic absorbance (2.2.25) : 360 to 390, determined at the
absorption maximum at 235 nm (anhydrous substance).
Dissolve 20.0 mg in water R and dilute to 100.0 mL with the
same solvent. Dilute 10.0 mL of the solution to 100.0 mL with
water R.
Related substances. Liquid chromatography (2.2.29). Prepare
the solutions immediately before use.
Solution A : mobile phase B, mobile phase A (14:86 V/V).
Test solution. Dissolve 40.0 mg of the substance to be
examined in solution A and dilute to 50.0 mL with the same
solution.
Reference solution (a). Dissolve 8.0 mg of cefotaxime acid CRS
in solution A and dilute to 10.0 mL with the same solution.
Reference solution (b). Dilute 1.0 mL of the test solution to
100.0 mL with solution A.
Reference solution (c). Add 1.0 mL of dilute hydrochloric acid R
to 4.0 mL of the test solution. Heat the solution at 40 C for
2 h. Add 5.0 mL of buffer solution pH 6.6 R and 1.0 mL of
dilute sodium hydroxide solution R.
Reference solution (d). Dissolve 4 mg of cefotaxime for peak
identification CRS (containing impurities A, B, C, E and F)
in 5 mL of solution A.
Column :
size : l = 0.15 m, = 3.9 mm,
stationary phase : octadecylsilyl silica gel for
chromatography R (5 m),
temperature : 30 C.
Mobile phase :
mobile phase A : 7.1 g/L solution of disodium hydrogen
phosphate R adjusted to pH 6.25 using phosphoric acid R ;
mobile phase B : methanol R ;
Time
(min)
0-7

Mobile phase A
(per cent V/V)
86

Mobile phase B
(per cent V/V)
14

7-9

86 82

14 18

9 - 16

82

18

16 - 45

82 60

18 40

45 - 50

60

40

50 - 55

60 86

40 14

55 - 60

86

14

System suitability : reference solution (c) :

resolution : minimum 3.5 between the peaks due to
impurity E and cefotaxime ;
symmetry factor : maximum 2.0 for the peak due to
cefotaxime.
Limits :
impurities A, B, C, D, E, F : for each impurity, not more
than the area of the principal peak in the chromatogram
obtained with reference solution (b) (1.0 per cent) ;
any other impurity : for each impurity, not more than
0.2 times the area of the principal peak in the chromatogram
obtained with reference solution (b) (0.2 per cent) ;
total : not more than 3 times the area of the principal peak
in the chromatogram obtained with reference solution (b)
(3.0 per cent) ;
disregard limit : 0.05 times the area of the principal peak
in the chromatogram obtained with reference solution (b)
(0.05 per cent).
Ethanol (2.4.24, System A) : maximum 1.0 per cent.
N,N-Dimethylaniline (2.4.26, Method B) : maximum 20 ppm.
2-Ethylhexanoic acid (2.4.28) : maximum 0.5 per cent m/m.
Water (2.5.12) : maximum 3.0 per cent, determined on 0.300 g.
Bacterial endotoxins (2.6.14) : less than 0.05 IU/mg, if
intended for use in the manufacture of parenteral preparations
without a further appropriate procedure for the removal of
bacterial endotoxins.
ASSAY
Liquid chromatography (2.2.29) as described in the test for
related substances with the following modication.
Injection : test solution and reference solution (a).
Calculate the percentage content of C16H16N5NaO7S2 by
multiplying the percentage content of cefotaxime by 1.048.
STORAGE
In an airtight container, protected from light. If the substance
is sterile, store in a sterile, airtight, tamper-proof container.
IMPURITIES
Specified impurities : A, B, C, D, E, F.
Other detectable impurities (the following substances would,
if present at a sufcient level, be detected by one or other of
the tests in the monograph. They are limited by the general
acceptance criterion for other/unspecied impurities and/or
by the general monograph Substances for pharmaceutical
use (2034). It is therefore not necessary to identify these
impurities for demonstration of compliance. See also 5.10.
Control of impurities in substances for pharmaceutical use) : G.

A. R = R = H : (6R,7R)-7-[[(2Z)-2-(2-aminothiazol-4Flow rate : 1.0 mL/min.

yl)-2-(methoxyimino)acetyl]amino]-3-methyl-8-oxoDetection : spectrophotometer at 235 nm.
5-thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylic acid
Injection : 10 L of the test solution and reference solutions (b),
(deacetoxycefotaxime),
(c) and (d).
B. R = OH, R = H : (6R,7R)-7-[[(2Z)-2-(2-aminothiazol-4Identification of impurities : use the chromatogram supplied
yl)-2-(methoxyimino)acetyl]amino]-3-(hydroxymethyl)-8with cefotaxime for peak identification CRS and the
oxo-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylic acid
chromatogram obtained with reference solution (d) to identify
(deacetylcefotaxime),
the peaks due to impurities A, B, C, E and F.
C. R = O-CO-CH3, R = CHO : (6R,7R)-3-[(acetyloxy)methyl]Relative retention with reference to cefotaxime (retention
time = about 13 min) : impurity B = about 0.3 ;
7-[[(2Z)-2-[2-(formylamino)thiazol-4-yl]-2impurity A = about 0.5 ; impurity E = about 0.6 ;
(methoxyimino)acetyl]amino]-8-oxo-5-thiaimpurity C = about 1.9 ; impurity D = about 2.3 ;
1-azabicyclo[4.2.0]oct-2-ene-2-carboxylic acid
impurity F = about 2.4 ; impurity G = about 3.1.
(N-formylcefotaxime),

1802

See the information section on general monographs (cover pages)

Cefoxitin sodium

EUROPEAN PHARMACOPOEIA 8.0

D. (6R,7R)-3-[(acetyloxy)methyl]-7-[[(2E)-2-(2aminothiazol-4-yl)-2-(methoxyimino)acetyl]amino]-8oxo-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylic acid
(E-cefotaxime),

DEFINITION
Sodium (6R,7S)-3-[(carbamoyloxy)methyl]-7-methoxy8-oxo-7-[[(thiophen-2-yl)acetyl]amino]-5-thia-1azabicyclo[4.2.0]oct-2-ene-2-carboxylate.
Semi-synthetic product derived from a fermentation product.
Content : 95.0 per cent to 102.0 per cent (anhydrous substance).
CHARACTERS
Appearance : white or almost white, very hygroscopic powder.
Solubility : very soluble in water, sparingly soluble in ethanol
(96 per cent).
IDENTIFICATION
A. Infrared absorption spectrophotometry (2.2.24).
Comparison : cefoxitin sodium CRS.
B. It gives reaction (a) of sodium (2.3.1).

E. (5aR,6R)-6-[[(2Z)-2-(2-aminothiazol-4-yl)-2(methoxyimino)acetyl]amino]-5a,6-dihydro-3H,7Hazeto[2,1-b]furo[3,4-d][1,3]thiazine-1,7(4H)-dione
(deacetylcefotaxime lactone),

TESTS
Solution S. Dissolve 2.50 g in carbon dioxide-free water R and
dilute to 25 mL with the same solvent.
Appearance of solution. Solution S is clear (2.2.1) and not
more intensely coloured than intensity 5 of the range of
reference solutions of the most appropriate colour (2.2.2,
Method II).
pH (2.2.3) : 4.2 to 7.0.
Dilute 2 mL of solution S to 20 mL with carbon dioxide-free
water R.
Specic optical rotation (2.2.7) : + 206 to + 214 (anhydrous
substance).
Dissolve 0.250 g in methanol R and dilute to 25.0 mL with
F. (6R,7R)-3-[(acetyloxy)methyl]-7-[[(2Z)-2-[2the same solvent.
[[[(6R,7R)-7-[[(2Z)-2-(2-aminothiazol-4-yl)-2Related substances. Liquid chromatography (2.2.29). Prepare
(methoxyimino)acetyl]amino]-2-carboxy-8-oxo-5-thiathe solutions immediately before use.
1-azabicyclo[4.2.0]oct-2-en-2-yl]methyl]amino]thiazolSolution A. Dissolve 1.0 g of potassium dihydrogen phosphate R
4-yl]-2-(methoxyimino)acetyl]amino]-8-oxo-5-thia-1and 1.8 g of anhydrous disodium hydrogen phosphate R in
azabicyclo[4.2.0]oct-2-ene-2-carboxylic acid (cefotaxime
1000 mL of water R. To 100 mL of the solution add 800 mL of
dimer),
water R, adjust to pH 7.0 with phosphoric acid R or a 40 g/L
solution of sodium hydroxide R and dilute to 1000 mL with
water R.
Test solution. Dissolve 50 mg of the substance to be examined
in solution A and dilute to 50.0 mL with solution A.
Reference solution (a). Dilute 1.0 mL of the test solution to
100.0 mL with solution A.
Reference solution (b). Dilute 1.0 mL of reference solution (a)
to 20.0 mL with solution A.
Reference solution (c). Dissolve 5 mg of cefoxitin for peak
identification CRS (containing impurities A, B, E, H, I and J)
G. (6R,7R)-3-[(acetyloxy)methyl]-7-[[(2Z)-2-[2-[[(2Z)in solution A and dilute to 5 mL with solution A.
2-(2-aminothiazol-4-yl)-2-(methoxyimino)aceColumn :
tyl]amino]thiazol-4-yl]-2-(methoxyimino)acetyl]ami size : l = 0.25 m, = 4.6 mm ;
no]-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylic acid (ATA cefotaxime).
stationary phase : phenylsilyl silica gel for chromatography R
(3.0 m) ;
temperature : 35 C.
01/2013:0990 Mobile phase :
mobile phase A : 1.0 g/L solution of ammonium formate R
CEFOXITIN SODIUM
adjusted to pH 2.7 with anhydrous formic acid R ;
mobile phase B : acetonitrile R ;

Cefoxitinum natricum

C16H16N3NaO7S2
[33564-30-6]

Time
(min)

Mobile phase A
(per cent V/V)

Mobile phase B
(per cent V/V)

0-5

92

5 - 50

92 74

8 26

50 - 85

74

26

Mr 449.4 Flow rate : 1.0 mL/min.

Detection : spectrophotometer at 254 nm.

General Notices (1) apply to all monographs and other texts

1803